RESUMO
AIMS: Fermented feed is an agricultural practice used in many regions of the world to improve the growth performance of farm animals. This study aimed to identify and evaluate the lactic acid bacteria and yeast involved in the production of fermented feed. METHODS AND RESULTS: We isolated and described two micro-organisms from autochthonous microbiota origin present in a regional feed product, Lactobacillus paracasei IBR07 (Lacticaseibacillus paracasei) and Kazachstania unispora IBR014 (Saccharomyces unisporum). Genome sequence analyses were performed to characterize both micro-organisms. Potential pathways involved in the acid response, tolerance and persistence were predicted in both genomes. Although L. paracasei and K. unispora are considered safe for animal feed, we analysed the presence of virulence factors, antibiotic resistance and pathogenicity islands. Furthermore, the Galleria mellonella model was used to support the safety of both isolates. CONCLUSIONS: We conclude that IBR07 and IBR014 strains are good candidates to be used as starter cultures for feed fermentation. SIGNIFICANCE AND IMPACT OF THE STUDY: The data presented here will be helpful to explore other biotechnological aspects and constitute a starting point for further studies to establish the consumption benefit of fermented feed in farm animal production.
Assuntos
Lacticaseibacillus paracasei , Lactobacillales , Ração Animal , Animais , Fermentação , Microbiologia de Alimentos , GenômicaRESUMO
The members of the Enterococcus genus are widely distributed in nature. Its strains have been extensively reported to be present in plant surfaces, soil, water and food. In an attempt to assess their potential application in food industry, four Enterococcus faecium group-strains recently isolated from Argentinean regional cheese products were evaluated using a combination of whole genome analyses and in vivo assays. In order to identify these microorganisms at species level, in silico analyses using their newly reported sequences were conducted. The average nucleotide identity (ANI), in silico DNA-DNA hybridization, and phylogenomic trees constructed using core genome data allowed IQ110, GM70 and GM75 strains to be classified as E. faecium while IQ23 strain was identified as E. durans. Besides their common origin, the strains showed differences in their genetic structure and mobile genetic element content. Furthermore, it was possible to determine the absence or presence of specific features related to growth in milk, cheese ripening, probiotic capability and gut adaptation including sugar, amino acid, and peptides utilization, flavor compound production, bile salt tolerance as well as biogenic amine production. Remarkably, all strains encoded for peptide permeases, maltose utilization, bile salt tolerance, diacetyl and tyramine production genes. On the other hand, some variability was observed regarding citrate and lactose utilization, esterase, and cell wall-associated proteinase. In addition, while strains were predicted to be non-human pathogens by the in silico inspection of pathogenicity and virulence factors, only the GM70 strain proved to be non-virulent in Galleria mellonella model. In conclusion, we propose that, in order to improve the rational selection of strains for industrial applications, a holistic approach involving a comparative genomic analysis of positive and negative features as well as in vivo evaluation of virulence behavior should be performed.
Assuntos
Queijo/microbiologia , Enterococcus faecium/classificação , Enterococcus faecium/genética , Inocuidade dos Alimentos/métodos , Genoma Bacteriano/genética , Animais , Argentina , Citratos/metabolismo , Enterococcus faecium/efeitos dos fármacos , Enterococcus faecium/isolamento & purificação , Esterases/genética , Sequências Repetitivas Dispersas/genética , Lactose/metabolismo , Maltose/metabolismo , Testes de Sensibilidade Microbiana , Leite/microbiologia , Tipagem Molecular/métodos , Mariposas/microbiologia , Probióticos , Virulência/genética , Fatores de Virulência/genéticaRESUMO
Bacillus pumilus group strains have been studied due their agronomic, biotechnological or pharmaceutical potential. Classifying strains of this taxonomic group at species level is a challenging procedure since it is composed of seven species that share among them over 99.5% of 16S rRNA gene identity. In this study, first, a whole-genome in silico approach was used to accurately demarcate B. pumilus group strains, as a case of highly phylogenetically related taxa, at the species level. In order to achieve that and consequently to validate or correct taxonomic identities of genomes in public databases, an average nucleotide identity correlation, a core-based phylogenomic and a gene function repertory analyses were performed. Eventually, more than 50% such genomes were found to be misclassified. Hierarchical clustering of gene functional repertoires was also used to infer ecotypes among B. pumilus group species. Furthermore, for the first time the machine-learning algorithm Random Forest was used to rank genes in order of their importance for species classification. We found that ybbP, a gene involved in the synthesis of cyclic di-AMP, was the most important gene for accurately predicting species identity among B. pumilus group strains. Finally, principal component analysis was used to classify strains based on the distances between their ybbP genes. The methodologies described could be utilized more broadly to identify other highly phylogenetically related species in metagenomic or epidemiological assessments.
RESUMO
We report the draft genome sequences of four Enterococcus faecium strains isolated from Argentine regional cheeses. These strains were selected based on their technological properties, i.e., their ability to produce aroma compounds (diacetyl, acetoin, and 2,3-butanediol) from citrate. The goal of our study is to provide further genetic evidence for the rational selection of enterococci strains based on their pheno- and genotype in order to be used in cheese production.
RESUMO
We report the draft genome sequence of Enterococcus mundtii CRL1656, which was isolated from the stripping milk of a clinically healthy adult Holstein dairy cow from a dairy farm of the northwestern region of Tucumán (Argentina). The 3.10-Mb genome sequence consists of 450 large contigs and contains 2,741 predicted protein-coding genes.
Assuntos
Enterococcus/classificação , Enterococcus/genética , Genoma Bacteriano , Animais , Argentina/epidemiologia , Bovinos , Feminino , Mastite Bovina/epidemiologia , Mastite Bovina/microbiologia , Leite/microbiologia , Dados de Sequência MolecularRESUMO
Over the last years an expanding family of small non-coding RNAs (sRNA) has been identified in eukaryotic genomes which behave as sequence-specific triggers for mRNA degradation, translation repression, heterochromatin formation and genome stability. To achieve their effectors functions, sRNAs associate with members of the Argonaute protein family. Argonaute proteins are segregated into three paralogous groups: the AGO-like subfamily, the PIWI-like subfamily, and the WAGO subfamily (for Worm specific AGO). Detailed phylogenetic analysis of the small RNA-related machinery components revealed that they can be traced back to the common ancestor of eukaryotes. However, this machinery seems to be lost or excessively simplified in some unicellular organisms such as Saccharomyces cerevisiae, Trypanosoma cruzi, Leishmania major and Plasmodium falciparum which are unable to utilize dsRNA to trigger degradation of target RNAs. We reported here a unique ORF encoding for an AGO/PIWI protein in T. cruzi which was expressed in all stages of its life cycle at the transcript as well as the protein level. Database search for remote homologues, revealed the presence of a divergent PAZ domain adjacent to the well supported PIWI domain. Our results strongly suggested that this unique AGO/PIWI protein from T. cruzi is a canonical Argonaute in terms of its domain architecture. We propose to reclassify all Argonaute members from trypanosomatids as a distinctive phylogenetic group representing a new subfamily of Argonaute proteins and propose the generic designation of AGO/PIWI-tryp to identify them. Inside the Trypanosomatid-specific node, AGO/PIWI-tryps were clearly segregated into two paralog groups designated as AGO-tryp and PIWI-tryp according to the presence or absence of a functional link with RNAi-related phenomena, respectively.
Assuntos
Proteínas de Protozoários/análise , Trypanosoma cruzi/classificação , Trypanosoma cruzi/metabolismo , Sequência de Aminoácidos , Animais , Clonagem Molecular , Evolução Molecular , Modelos Moleculares , Dados de Sequência Molecular , Filogenia , Proteínas de Protozoários/química , RNA Interferente Pequeno , Homologia de Sequência do Ácido NucleicoRESUMO
Recent findings associate transcription start in trypanosomatids with chromatin regions containing modified and variant histones. TATA-binding protein (TBP) and other fundamental transcription factors have been also found at these Transcription Start Sites (TSS). Results of Systematic Evolution of Ligands by Exponential Enrichment (SELEX) experiments show that Trypanosoma cruzi TBP (TcTBP) has an in vitro binding preference for G-rich sequences. This finding correlates with the presence of G-rich stretches at the Strand Switch Regions (SSR) and at some putative internal TSS in Trypanosoma brucei and Leishmania. A scanning study of partially assembled T. cruzi genomic contigs determined the presence of G-rich stretches in the coding strands. TcTBP affinity for G-rich sequences suggests that this factor could play a role in locating the initiation complex in the right TSS, probably by "sensing" the G-content on the strand to be transcribed.
Assuntos
Sequência Rica em GC/fisiologia , Proteína de Ligação a TATA-Box/metabolismo , Trypanosoma cruzi/metabolismo , Sequência Consenso , Trypanosoma cruzi/genéticaRESUMO
El monofluorfosfato de sodio (MFP) es una droga que se prescribe como fuente de fluoruro para el tratamiento de la osteoporosis. El fluoruro es la especie mitogenica que estimula la diferenciacion y proliferacion de los osteoblastos. Posee mejor tolerancia gastrica que el NaF y se puede administrar con suplementos de calcio. Empiricamente se lo ha utilizado como equivalente del NaF aunque exhibe sustanciales diferencias farmacocineticas. Luego de una dosis oral del MFP, una pequeña fraccion de la misma se absorbe intacta y se liga a algunas globulinas del plasma formando un compartimiento de fluor no difusible (fluor ligado a proteinas), que no se observa cunado se administra NaF. El ion fluoruro no se liga a las proteinas del plasma. Cuando se comparan las curvas de fluoremia total en funcion del tiempo despues de la administracion de dosis equimolares de MFP y NaF se comprueba, en animales de experimentacion y en voluntarios humanos, que el area bajo la curva de MFP duplica la de NaF. Estos reaultados explican la reduccion a la mitad de la dosis terapeutica de fluor cuando se emplea MFP. El objetivo central de esta tesis fue develar el metabolismo del MFP para explicar la mayor tolerancia y eficiencia terapeutica, relacionadas con la mayor biodisponibilidad de fluor (respecto del NaF usado como standard). Accesoriamente surgio como un objetivo secundario identificar las caracteristicas de ligamiento de MFP a alfa2-M. La primera etapa de la investigacion se oriento a identificar las proteinas responsables de la aparicion del comportamiento de fluor ligado a proteinas. El MFP se liga principalmente a la alfa-globulinas y en menor grado a las beta-glñobulinas. El complejo MFP-globulina es estable y resiste cambios de pH, efecto de quelantes, dilucion o hidrolisis del MFP por fosfatasa alcalina. El ligamiento de MFP a las globulinas se aparta del modelo descripto por Scatchard de n sitios independientes y equivalentes. Los resultados experimentales sugieren un modelo de cooperativismo positivo. La alfa 2-macroglobulina (alfa2-M) es la proteina principalmente responsable de la fijacion de MFP. Al formarse el complejo MFP-alfa2-M, la proteina pierde su actividad como antiproteasa. El componente C3 del complemento, que comparte semejanzas estrurales con alfa2-M tambien liga MFP perdiendo su actividad biologica. La formacion del complejo MFP-proteinas con la concurrenteperdida de la actividad biologica de las proteinas participantes...
Assuntos
Humanos , Idoso , Ratos , Animais , Técnicas In Vitro , Macroglobulinas , Osteoporose , Fluoreto de SódioRESUMO
El monofluorfosfato de sodio (MFP) es una droga que se prescribe como fuente de fluoruro para el tratamiento de la osteoporosis. El fluoruro es la especie mitogenica que estimula la diferenciacion y proliferacion de los osteoblastos. Posee mejor tolerancia gastrica que el NaF y se puede administrar con suplementos de calcio. Empiricamente se lo ha utilizado como equivalente del NaF aunque exhibe sustanciales diferencias farmacocineticas. Luego de una dosis oral del MFP, una pequeña fraccion de la misma se absorbe intacta y se liga a algunas globulinas del plasma formando un compartimiento de fluor no difusible (fluor ligado a proteinas), que no se observa cunado se administra NaF. El ion fluoruro no se liga a las proteinas del plasma. Cuando se comparan las curvas de fluoremia total en funcion del tiempo despues de la administracion de dosis equimolares de MFP y NaF se comprueba, en animales de experimentacion y en voluntarios humanos, que el area bajo la curva de MFP duplica la de NaF. Estos reaultados explican la reduccion a la mitad de la dosis terapeutica de fluor cuando se emplea MFP. El objetivo central de esta tesis fue develar el metabolismo del MFP para explicar la mayor tolerancia y eficiencia terapeutica, relacionadas con la mayor biodisponibilidad de fluor (respecto del NaF usado como standard). Accesoriamente surgio como un objetivo secundario identificar las caracteristicas de ligamiento de MFP a alfa2-M. La primera etapa de la investigacion se oriento a identificar las proteinas responsables de la aparicion del comportamiento de fluor ligado a proteinas. El MFP se liga principalmente a la alfa-globulinas y en menor grado a las beta-glñobulinas. El complejo MFP-globulina es estable y resiste cambios de pH, efecto de quelantes, dilucion o hidrolisis del MFP por fosfatasa alcalina. El ligamiento de MFP a las globulinas se aparta del modelo descripto por Scatchard de n sitios independientes y equivalentes. Los resultados experimentales sugieren un modelo de cooperativismo positivo. La alfa 2-macroglobulina (alfa2-M) es la proteina principalmente responsable de la fijacion de MFP. Al formarse el complejo MFP-alfa2-M, la proteina pierde su actividad como antiproteasa. El componente C3 del complemento, que comparte semejanzas estrurales con alfa2-M tambien liga MFP perdiendo su actividad biologica. La formacion del complejo MFP-proteinas con la concurrenteperdida de la actividad biologica de las proteinas participantes...(AU)
Assuntos
Humanos , Idoso , Ratos , Animais , Técnicas In Vitro , Macroglobulinas/uso terapêutico , Fluoreto de Sódio , OsteoporoseRESUMO
Sodium monofluorophosphate (MFP) is a drug used in the treatment of primary osteoporosis. Following the intake of MFP, a small fraction of the drug is absorbed intact and forms a complex with alpha2-macroglobulin (MFP-alpha2M) inactivating the antiproteasic activity of the globulin. The complex has been shown to occur in the serum of rats and human being. This paper reports data on the metabolism of this complex in the rat. In vitro experiments showed that liver and bone tissue remove MFP-alpha2M from the incubation medium. When the experiments were pursued beyond the time needed to reduce the complex concentration to very low levels, fluorine (F) reappears in the medium in two forms: bound to low molecular weight macromolecule/s (2,200 + 600 Da) and as ionic F. Concentrations of these F fractions increase while that of the complex decreases as a function of time. In vitro, uptake of the complex by liver or bone tissue was not affected by the presence of colchicine or methylamine. These drugs, however, inhibited intracellular metabolism of the complex, as indicated by the impairment of the return of F species to the extracellular space and the increase in F content of the tissue. The cellular receptors responsible for the uptake of the complex in liver and bone are insensitive to low concentration of calcium and inhibited by polyinosinic acid[5']. These features characterize the "scavenger" receptor, one of the two receptor types known to remove inactive alpha2M from the circulation. Injection of polyinosinic acid [5'] to living rats also hindered the disappearance of the complex from serum. It is concluded that the metabolism of the MFP-alpha2M complex involves binding to receptors, uptake by cells, lysosomal degradation and return of F bound to low molecular weight macromolecule/s to the extracelular space. It is assumed, however, that inorganic F is the final product of lysosomal hydrolysis of the protein moiety.
Assuntos
Animais , Feminino , Ratos , alfa-Macroglobulinas/farmacocinética , Osso e Ossos/efeitos dos fármacos , Fígado/efeitos dos fármacos , Fosfatos/farmacocinética , Disponibilidade Biológica , Flúor/análise , Flúor/metabolismo , Substâncias Macromoleculares , Peso MolecularRESUMO
Sodium monofluorophosphate (MFP) is a drug used in the treatment of primary osteoporosis. Following the intake of MFP, a small fraction of the drug is absorbed intact and forms a complex with alpha2-macroglobulin (MFP-alpha2M) inactivating the antiproteasic activity of the globulin. The complex has been shown to occur in the serum of rats and human being. This paper reports data on the metabolism of this complex in the rat. In vitro experiments showed that liver and bone tissue remove MFP-alpha2M from the incubation medium. When the experiments were pursued beyond the time needed to reduce the complex concentration to very low levels, fluorine (F) reappears in the medium in two forms: bound to low molecular weight macromolecule/s (2,200 + 600 Da) and as ionic F. Concentrations of these F fractions increase while that of the complex decreases as a function of time. In vitro, uptake of the complex by liver or bone tissue was not affected by the presence of colchicine or methylamine. These drugs, however, inhibited intracellular metabolism of the complex, as indicated by the impairment of the return of F species to the extracellular space and the increase in F content of the tissue. The cellular receptors responsible for the uptake of the complex in liver and bone are insensitive to low concentration of calcium and inhibited by polyinosinic acid[5]. These features characterize the "scavenger" receptor, one of the two receptor types known to remove inactive alpha2M from the circulation. Injection of polyinosinic acid [5] to living rats also hindered the disappearance of the complex from serum. It is concluded that the metabolism of the MFP-alpha2M complex involves binding to receptors, uptake by cells, lysosomal degradation and return of F bound to low molecular weight macromolecule/s to the extracelular space. It is assumed, however, that inorganic F is the final product of lysosomal hydrolysis of the protein moiety. (AU)
Assuntos
Animais , Feminino , Ratos , Fígado/efeitos dos fármacos , Osso e Ossos/efeitos dos fármacos , alfa-Macroglobulinas/farmacocinética , Fosfatos/farmacocinética , Peso Molecular , Substâncias Macromoleculares , Flúor/análise , Flúor/metabolismo , Disponibilidade BiológicaRESUMO
El autor describe paso a paso la técnica de reconstrucción del ligamento cruzado anterior de la rodilla por vía transtendón rotuliano, utilizando como sustituto el tercio central del tendón patelar como alternativa al uso de la asistencia artroscópica cuando económicamente no es posible
Assuntos
Ligamento Cruzado Anterior/cirurgia , ArgentinaRESUMO
El autor describe paso a paso la técnica de reconstrucción del ligamento cruzado anterior de la rodilla por vía transtendón rotuliano, utilizando como sustituto el tercio central del tendón patelar como alternativa al uso de la asistencia artroscópica cuando económicamente no es posible
Assuntos
Argentina , Ligamento Cruzado Anterior/cirurgiaRESUMO
El autor propone analizar cuál es la conducta terapéutica más conveniente frente a una lesión aguda o crónica de rodilla, donde se ven afectados en grado variable los elementos que proveen la estabilidad. Para ello analiza cuáles son estos elementos y cuál la definición de estabilidad, estática y dinámica. Luego realiza precisiones sobre lo que a su criterio deben ser rasgos terapéuticos aceptados frente a una lesión de rodilla, descartando la idea de reparar solamente el ligamento cruzado anterior, encarando en cambio la reparación/reconstrucción anatómica de todos los elementos dañados, incluyendo los ligamentos laterales. Hace una reseña de los casos tratados en entes oficiales y privados en los últimos 35 años y analiza los resultados, siguiendo los Criterios de Toronto. Concluye diciendo que frente a una inestabilidad compleja de rodilla no alcanza con reparar o reconstruir solamente el ligamento cruzado anterior, sino que debe encararse, en uno o más tiempos, la reparación/reconstrucción anatómica de todos los elementos dañados
Assuntos
Joelho , Ferimentos e Lesões , Ligamentos , Argentina , Instabilidade ArticularRESUMO
El autor propone analizar cuál es la conducta terapéutica más conveniente frente a una lesión aguda o crónica de rodilla, donde se ven afectados en grado variable los elementos que proveen la estabilidad. Para ello analiza cuáles son estos elementos y cuál la definición de estabilidad, estática y dinámica. Luego realiza precisiones sobre lo que a su criterio deben ser rasgos terapéuticos aceptados frente a una lesión de rodilla, descartando la idea de reparar solamente el ligamento cruzado anterior, encarando en cambio la reparación/reconstrucción anatómica de todos los elementos dañados, incluyendo los ligamentos laterales. Hace una reseña de los casos tratados en entes oficiales y privados en los últimos 35 años y analiza los resultados, siguiendo los Criterios de Toronto. Concluye diciendo que frente a una inestabilidad compleja de rodilla no alcanza con reparar o reconstruir solamente el ligamento cruzado anterior, sino que debe encararse, en uno o más tiempos, la reparación/reconstrucción anatómica de todos los elementos dañados